Figure 3. Histomorphometry methods.
Safranin O stained histological cross sections (A) were manually segmented in Photoshop to create a mask indicating the location of the tissue boundary (black border around tissue) and the lacunae (black circles) within the tissue (B). Masks were then analyzed with a custom Matlab program to divide the tissue into five equal regions (C) and calculate the number of cells and the area occupied by cells within each region.